Optimized time-gated generalized polarization imaging of Laurdan and di-4-ANEPPDHQ for membrane order image contrast enhancement
✍ Scribed by Dylan M. Owen; Katharina Gaus
- Publisher
- John Wiley and Sons
- Year
- 2009
- Tongue
- English
- Weight
- 394 KB
- Volume
- 73
- Category
- Article
- ISSN
- 1059-910X
No coin nor oath required. For personal study only.
✦ Synopsis
Abstract
The membrane dyes Laurdan and di‐4‐ANEPPDHQ can be used to image membrane order due to a spectral blue‐shift in the fluorescence emission between the liquid‐ordered and liquid‐disordered phases. These images typically take the form of a normalized intensity ratio image known as a generalized polarization (GP) plot. Here, we exploit the known excited state photophysics and time‐resolved data acquisition via time‐correlated single‐photon counting (TCSPC) to demonstrate GP contrast enhancement for these two probes of 7 and 31%, respectively. This improvement in image contrast enhancement will be invaluable when studying the role of lipid rafts in fixed and live cell systems. Microsc. Res. Tech. 2010. © 2009 Wiley‐Liss, Inc.